Disease Information(sort of like the Intro to your MiniResearch Writeup):
Malaria is a common infectious disease caused by the protozoal parasite, Plasmodium vivax. This parasite is carried by mosquitos popular to low and middle income countries. The disease begins with an infection of the red blood cells leading to fever. Infected blood cells can also become plugged in the capillaries, depriving tissues of oxygen. The parasite can continue to multiply and cause more serious symptoms including seizures, coma, or death. It is difficult to diagnose malaria because the parasite, Plasmodium vivax, can live dormant in the liver for months to years, causing no symptoms and not showing up in blood tests. A single bite from an infected mosquito can cause six or more relapses of the disease in a year. Chloroquine is currently the drug treatment of choice, however, people are becoming increasingly chloroquine resistant which is causing problems in infectious breakouts.
DNA polymerase alpha is an enzyme in bacteria because it catalyzes DNA template directed extension of the 3’ end of a strand of DNA one nucleotide at a time. Inhibition of this enzyme would stop the replication of DNA inside of the parasite Plasmodium vivax, therefore killing it.
Is it a monomer or multimer as biological unit: dimer
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
The druggable index for DNA polymerase alpha on 0-1 scale is a 0.8. This target has been proved to be druggable in many other organisms, one including hepatitis C.
Tedesco, R.; Bambal, R.; Chai, D.; Dhanak, D.; 3-(1,1-dioxo-2H-(1,2,4)-benzothiadiazin-3-yl)-4-hydroxy-2(1H)-quinolinones, potent inhibitors of hepatitis C virus RNA-dependent RNA polymerase. Journal of Medical Chemistry 2006, 49 (3) 971-83.
*CDSGene Sequence(paste as text only): >5IJL:A|PDBID|CHAIN|SEQUENCE GTGDGSELPKEMEEYFEMLQREIDKAYEIAKKARAQGKDPSLDVEIPQATDMAGRVESLVGPPGVAKRIRELVKEYGKEI AALKIVDEIIEGKFGDLGSREKYAEQAVRTALAILTEGIVSAPIEGIANVKIKRNTWADNSEYLALYYAGPIRSSGGTAQ ALSVLVGDYVRRKLGLDRFKPSEKHIERMVEEVDLYHRAVTRLQYHPSPEEVRLAMRNIPIEITGEATDDVEVSHRDVPG VETNQLRGGAILVLAEGVLQKAKKLVKYIDKMGIEGWEWLKEFVEAKEKGEPKEEGKEESLAESTLEETKVEVDMGFYYSLYQKFKEEIAPSDKYAKEVIGGRPLFSDPSKPGGFRLRYGRSRASGFATWGINPATMILVDEFLAIGTQLKTERPGKGAV VTPVTTIEGPIVKLKDGSVLRVDDYNLALKVREDVEEILYLGDAVIAFGDFVENNQTLLPANYCEEWWILEFVKALKEIY EVHLEPFTENEEESIEEASDYLEIDPEFLKEMLRDPLRVKPPVELAIHFSEVLGIPLHPYYTLYWNSVEPKDVEKLWRLL KNYAEIEWSNFRGIKFAKKIVISQEKLGDSKRTLELLGLPHTVRDGNVIVDYPWAAALLTPLGNLNWEFMAKPLYATIDI INENNEIKLRDRGISWIGARMGRPEKAKERKMKPPVQVLFPIGLAGGSSRDIKKAAEEGKVAEVEIAFFKCPKCGHVGPE HLCPNCGTRKELLWVCPRCNAEYPESQAEGYNYTCPKCNVKLRPYAKRKIRPSELLNRAMENVKVYGVDKLKGVMGMTSGWKMPEPLEKGLLRAKNDVYVFKDGTIRFDATDAPITHFRPREIGVSVEKLRELGYTHDFEGKPLVSEDQIVELKPQDIILSKEAGRYLLKVAKFVDDLLEKFYGLPRFYNAEKMEDLIGHLVIGLAPHTSAGIVGRIIGFVDALVGYAHPYFHAAKRRNC DGDEDAVMLLLDALLNFSRYYLPEKRGGKMDAPLVITTRLDPREVDSEVHNMDIVRYYPLEFYEATYELKSPKELVGVIE RVEDRLGKPEMYYGLKFTHDTDDIAL
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
*GC% Content for gene: output
*GC% Content for gene (codon optimized):
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works outputtext file-that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
Target (protein/gene name):DNA polymerase alpha, positive
NCBI #:EDL43576.1
Protein ID (NP or XP #) or Wolbachia#:27548043
Organism (including strain):malaria parasite Plasmodium vivax
Etiologic Risk Group (see link below):RG2
Disease Information(sort of like the Intro to your MiniResearch Writeup):
Malaria is a common infectious disease caused by the protozoal parasite, Plasmodium vivax. This parasite is carried by mosquitos popular to low and middle income countries. The disease begins with an infection of the red blood cells leading to fever. Infected blood cells can also become plugged in the capillaries, depriving tissues of oxygen. The parasite can continue to multiply and cause more serious symptoms including seizures, coma, or death. It is difficult to diagnose malaria because the parasite, Plasmodium vivax, can live dormant in the liver for months to years, causing no symptoms and not showing up in blood tests. A single bite from an infected mosquito can cause six or more relapses of the disease in a year. Chloroquine is currently the drug treatment of choice, however, people are becoming increasingly chloroquine resistant which is causing problems in infectious breakouts.
Link to TDR Targets page (if present):http://tdrtargets.org/targets/view?gene_id=265629
Link to Gene Database page NCBI:https://www.ncbi.nlm.nih.gov/protein/EDL43576.1
Essentiality of this protein:
DNA polymerase alpha is an enzyme in bacteria because it catalyzes DNA template directed extension of the 3’ end of a strand of DNA one nucleotide at a time. Inhibition of this enzyme would stop the replication of DNA inside of the parasite Plasmodium vivax, therefore killing it.
Is it a monomer or multimer as biological unit: dimer
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
The druggable index for DNA polymerase alpha on 0-1 scale is a 0.8. This target has been proved to be druggable in many other organisms, one including hepatitis C.
Tedesco, R.; Bambal, R.; Chai, D.; Dhanak, D.; 3-(1,1-dioxo-2H-(1,2,4)-benzothiadiazin-3-yl)-4-hydroxy-2(1H)-quinolinones, potent inhibitors of hepatitis C virus RNA-dependent RNA polymerase. Journal of Medical Chemistry 2006, 49 (3) 971-83.
EC#:2.7.7.7
Link to BRENDA EC# page:http://www.brenda-enzymes.org/enzyme.php?ecno=2.7.7.7
Enzyme Assay information:
http://www.sigmaaldrich.com/catalog/product/roche/taqro?lang=en®ion=US
Structure (PDB or Homology model)
PDB #: 5IJL
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
ORIGIN
1 msnvfdklkk qrskeckatd fyevknkndd iyevvdeegy ienknslknf ivggdkdfds
61 sddvivedin ygllrkrtae keakakrntp iddfirekki qkifstgkls keeiskakdi
121 erlmdansss sgsesdhfna krktrdsyer drssregchi rqivrggrae gqvktqvkae
181 heenvdataa dvdegdldge dhldgegtpe rskkrrvltg gatpegvkyt pgdfsteick
241 geeedvkkrn ergtplcgik qestpegqss dqrsivarle enhpvetflq mgdpppatik
301 mknevrgspc dgeddqeksr littigmknk pdelvtneet gfvnvyifdi ckhrnsiimf
361 grtltkfkiy rsvsiyiedm yrcyyfllsk tmvykkdgve ikyghknynt cimsdfleef
421 ravrkyhnie sikckivkrr nlnlsssded vyakifytyn cdpiseryls grtysafycc
481 nedvienfvi kknlklpcwv kvrglklnlt nnltycyfdc ttanwknvyp vekrlggkaa
541 iggvsavsgg ssggsaangg saanggstpn gvaaeeghrk gdpsaggsgh ladldlnkvv
601 ikvvsllnee nhhevfsics mvdaegmkyi nfcgistkaa kkknltynks naklfdsekn
661 lldaflqklk tldtdiyigy nilnfdlefv ihrcnvhnln adflsrkkkl kkndkmkvnk
721 fngtsstgsm yniiqnikgr livdiymlck dlikltsygm deivdsvrgr fqegasaaka
781 anaanaasaa rssnkaskan aasgtastss tasassppnv fkdfsvnnin llncsnihly
841 daqqllenhl nmyihsqlfc vqeianvckv lqiiektrdl tklsgylwar sllcytseri
901 eyfllheynr kkfitpvvkk rvkkidseqv knkaaakysg glvleplcgy ydtfvlyldf
961 nslypsiiie ynvcfstlkg rgeaanlgea anlgeaanrg eaanlgeaan lgeaanlgai
1021 stelgdaead aavalseave vddfdrtkpg ilpsilknlv ekrafikkli aneknkekke
1081 llliqslsik lisnsiygcl gnsnnrfyak hmaafitlkg rnllqhtksr verefnlkvi
1141 ygdtdsimid tgikaqggsn ykeafrlahv ikntinksyk rleldlecif skllllkkkk
1201 yacakvvdan merceyemkg infikrdfsr isklignevl kiifsskdgq avgggggags
1261 agsadppven dlseqiheyl rgvhqrilad efdldfftit kkltknvsey qeknslghvl
1321 vaermmkdgy nisvnkeiqy cvctsedacr ffnkpghekl snslccfsls eihkhnlrvd
1381 kdyyirnqil spinrlcqyi egtsaeklas cfniydvrei ktdlqieeny qeanvlslln
1441 eseerfkdih lkgyltcsyc shnvkpaifi kyfrcnnclr ylsieqirny ifsflhhlfa
1501 tfykqlyvcs scmvktrrvl lngakdcpsp ncespkgslk plvskkyiyl mleyflfllk
1561 dnlrevppgq haantatapt taegaapyas dvlvyvcvde kfkthissdq kpgkknlrve
1621 ayddicnkke lgvtitkglq mkypnisnfi tylnlkssqf cinyneertt irhslqtlvr
1681 nnvyaqiyfd rvfsvfqqpl agkiaql
//
Length of your protein in Amino Acids: 1707 amino acids.
Molecular Weight of your protein in kiloDaltons: 192.17 kDA
Molar Extinction coefficient of your protein at 280 nm wavelength: 110380 mol^-1cm-1
TMpred graph Image (http://www.ch.embnet.org/cgi-bin/TMPRED_form_parser)
*CDS Gene Sequence (paste as text only):
>5IJL:A|PDBID|CHAIN|SEQUENCE
GTGDGSELPKEMEEYFEMLQREIDKAYEIAKKARAQGKDPSLDVEIPQATDMAGRVESLVGPPGVAKRIRELVKEYGKEI
AALKIVDEIIEGKFGDLGSREKYAEQAVRTALAILTEGIVSAPIEGIANVKIKRNTWADNSEYLALYYAGPIRSSGGTAQ
ALSVLVGDYVRRKLGLDRFKPSEKHIERMVEEVDLYHRAVTRLQYHPSPEEVRLAMRNIPIEITGEATDDVEVSHRDVPG
VETNQLRGGAILVLAEGVLQKAKKLVKYIDKMGIEGWEWLKEFVEAKEKGEPKEEGKEESLAESTLEETKVEVDMGFYYSLYQKFKEEIAPSDKYAKEVIGGRPLFSDPSKPGGFRLRYGRSRASGFATWGINPATMILVDEFLAIGTQLKTERPGKGAV
VTPVTTIEGPIVKLKDGSVLRVDDYNLALKVREDVEEILYLGDAVIAFGDFVENNQTLLPANYCEEWWILEFVKALKEIY
EVHLEPFTENEEESIEEASDYLEIDPEFLKEMLRDPLRVKPPVELAIHFSEVLGIPLHPYYTLYWNSVEPKDVEKLWRLL
KNYAEIEWSNFRGIKFAKKIVISQEKLGDSKRTLELLGLPHTVRDGNVIVDYPWAAALLTPLGNLNWEFMAKPLYATIDI
INENNEIKLRDRGISWIGARMGRPEKAKERKMKPPVQVLFPIGLAGGSSRDIKKAAEEGKVAEVEIAFFKCPKCGHVGPE
HLCPNCGTRKELLWVCPRCNAEYPESQAEGYNYTCPKCNVKLRPYAKRKIRPSELLNRAMENVKVYGVDKLKGVMGMTSGWKMPEPLEKGLLRAKNDVYVFKDGTIRFDATDAPITHFRPREIGVSVEKLRELGYTHDFEGKPLVSEDQIVELKPQDIILSKEAGRYLLKVAKFVDDLLEKFYGLPRFYNAEKMEDLIGHLVIGLAPHTSAGIVGRIIGFVDALVGYAHPYFHAAKRRNC
DGDEDAVMLLLDALLNFSRYYLPEKRGGKMDAPLVITTRLDPREVDSEVHNMDIVRYYPLEFYEATYELKSPKELVGVIE
RVEDRLGKPEMYYGLKFTHDTDDIAL
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
*GC% Content for gene:
output
*GC% Content for gene (codon optimized):
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
**